Isolation of Neurospora mitochondrial tRNA.
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Biomedical subjects
Publications and source records attributed to W E Barnett.
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Fragments of Euglena chloroplast DNA generated by endonuclease R-Eco RI were separated by agarose-gel electrophoresis into 24 distinct bands. At least five fragments contain sequences complementary to chloroplast ribosomal RNA, Most of the Eco RI fragments have been cloned in a plasmid of Escherichia coli. Three of the cloned fragments were shown to contain chloroplast ribosomal RNA sequences by DNA-RNA hybridization.
Initiator methionine tRNA from the cytoplasm of Neurospora crassa has been purified and sequenced. The sequence is: pAGCUGCAUm1GGCGCAGCGGAAGCGCM22GCY*GGGCUCAUt6AACCCGGAGm7GU (or D) - CACUCGAUCGm1AAACGAG*UUGCAGCUACCAOH. Similar to initiator tRNAs from the cytoplasm of other eukaryotes, this tRNA also contains the sequence -AUCG- instead of the usual -TphiCG (or A)- found in loop IV of other tRNAs. The sequence of the N. crassa cytoplasmic initiator tRNA is quite different from that of the corresponding mitochondrial initiator tRNA. Comparison of the sequence of N. crassa cytoplasmic initiator tRNA to those of yeast, wheat germ and vertebrate cytoplasmic initiator tRNA indicates that the sequences of the two fungal tRNAs are no more similar to each other than they are to those of other initiator tRNAs.
A response to: "A consideration of Euglena gracilis W3BUL as a cytoplasmic control for the wild-type phenylalanyl-tRNA synthetase system" and "A reinvestigation of the sites of transcription and translation of Euglena chloroplastic phenylalanyl-tRNA synthetase" by J. L. Lesiewicz and D. S. Herson.
The primary sequence of phenylalanine tRNA (tRNAphe) from the chloroplasts of Euglena gracilis has been determined: pG-C-U-G-G-G-A-U-A-G-C-U-C-A-G-D-U-G-Gm-U(U)-A-G-A-G-C-G-G-A-G-G-A-C-U-G-A-A-A-A-PSI-C-C-U-U-G-U-m7G-Py-C-A-C-C-A-G-T-psi-C-A-A-A-U-C-U-G-G-U-U-C-C-U-A-G-C-A-C-C-A. This represents the first nucleotide sequence determined for an organelle tRNA. As do all other tRNAPhes thus far sequenced, chloroplastic tRNAPhe contains 76 nucleotides. Both in the nature of its modified nucleotides and in its sequence (although the sequence of all known tRNAPhes is quite similar), chloroplastic tRNAPhe more closely resembles procaryotic tRNAPhes than it resembles those from the cytoplasm of eucaryotes. There are eight positions in the tRNAPhe molecule where nucleotides are invariant in procaryotes but differ from invariant nucleotides in eucaryotes; at five of these positions, chloroplastic tRNAPhe is similar to procaryotes. The possible evolutionary significance of this intermediate type of structure is discussed.
The nucleoside compositions of chloroplast and cytoplasmic tRNAPhe's from Euglena gracilis have been determined. The modified nucleoside compositions of these two tRNAs indicate that tRNAPheChl is more similar to procaryotic (E. coli) tRNAPhe than to either the Euglena cytoplasmic tRNAPhe or other eucaryotic cytoplasmic tRNAPhe's.